Fig 2: LAIR-1 overexpression inhibits HOS cell migration. a Representative images of cell migration for wound closure. b Statistical ratio of wound closure after scratch formation. c Representative images of transwell membranes stained with crystal violet showing a decreased number of cells after LAIR-1 overexpression. d Relative ratio of migratory cells per field. e–g LAIR-1 overexpression changes the mRNA expression levels of EMT markers in HOS cells, as determined by qPCR. * P < 0.05

Fig 3: LAIR-1 overexpression in HOS cells inhibits the expression of EMT-associated transcription factors by decreasing p-Foxo1 expression. a Expression of Twist1 in untreated HOS cells (Blank) and corresponding LV-NC- and LV-LAIR-1-overexpressing transfectants was analyzed by qPCR. b Twist1, N-cadherin, p-Foxo1, total Foxo1, p-Akt, and total Akt expression were analyzed by western blotting in blank, LV-NC-overexpressing, and LV-LAIR-1-overexpressing OS cells. c Twist1, p-Foxo1, and total Foxo1 expression were analyzed by western blotting in blank, negative control (NC) siRNA, and Foxo1 siRNA (siFoxo1)-transfected OS cells. Representative immunofluorescence staining for Twist1 (d) and Foxo1 (e) in HOS cells (left). Cell nuclei were stained with DAPI (blue). Scale bar = 20 μm. The statistical data for the images are shown in histograms (right). *P < 0.05, **P < 0.01. Results represent at least three independent experiments.

Fig 4: LAIR-1 inhibits Glut1-related glucose uptake in OS cells. a Heatmap showing the levels of differentially expressed mRNAs. b Top 20 KEGG pathway annotation categories for target gene functions of predicted mRNAs. c Selected significantly differentially expressed mRNA-related to EMT in RNA-seq data between two groups, ***P < 0.001. d qPCR validation of differentially expressed EMT-related genes in LV-NC and LV-LAIR-1-overexpressing OS cells, **P < 0.01. e Glut1 expression analyzed by western blotting. f Immunofluorescence staining of Glut1 in the LV-LAIR-1-overexpressing OS cells. Scale bar = 50 μm. Data were obtained from at least two independent experiments.